SmartRT™ Reverse Transcriptase Kit

$0.00$338.77

SmartRTTM Reverse Transcriptase is an engineered MMLV RT that improves the thermostability of the enzyme, reduces RNase H activity and its cDNA synthesis ability. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized cDNA.  Together with a 3’ modified oligo dT primer and a 5’  SMART universal oligo, which contains a terminal complementation to nucleic acids at the 3’  end of the first-strand cDNA, the SmartRTTM reverse transcriptase produces RACE ready full-length cDNA.

SKU OPTIONS PriceQuantity
SMRT-100 4000u(40 rxn) (100u/ul) $55.52
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SMRT-200 10000u(100 rxn) (100u/ul) $100.84
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SMRT-300 40000u(400 rxn) (100u/ul) $338.77
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SMRT-OEM any size Please inquire
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Description

General Description:
SmartRTTM Reverse Transcriptase is an engineered MMLV RT that improves the thermostability of the enzyme, reduces RNase H activity and its cDNA synthesis ability. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized cDNA.  Together with a 3’ modified oligo dT primer and a 5’  SMART universal oligo, which contains a terminal complementation to nucleic acids at the 3’  end of the first-strand cDNA, the SmartRTTM reverse transcriptase produces RACE ready full-length cDNA.

Feature:
MCLAB’s reverse transcriptase is one of the transcriptases that has a terminal transferase activity with the highest thermostability.

Source:
expressed in E. coli

Application:
Reverse transcription and RACE ready full-length cDNA synthesis

Supplied with:
5 x first strand cDNA synthesis buffer

Recommended Storage Condition:
-20 °C

Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in 10 minutes at 37°C using poly(A):oligo(dT) as template:primer.

Shipping Condition:
Dry ice

User Protocol:
For RACE-ready first-strand cDNA Synthesis
Combine the following in separate 0.2-ml PCR tubes (20 µl reaction):
1–9 µl RNA sample
1 µl 5’ SMART universal Primer
1 µl oligo dT Primer
1 µl dNTP
Add sterile H2O to a final volume of 12 µl for each reaction.
Incubate the tubes at 65°C for 5 min
Cool the tubes on ice for 3 min
Add the following to each reaction tube:
4 µl 5X First-Strand Buffer
1 µl DTT (20 mM)
2 µl dNTP Mix (10 mM)
2 µl SmartRT reverse transcriptase
Incubate the tubes:
50°C for 1.5 hr
85°C for 5 min

For regular first-strand cDNA Synthesis:

Combine the following in separate 0.2-ml PCR tubes (20 µl reaction):
1–10 µl RNA sample
1 µl oligo dT Primer or Random primer
1 µl dNTP
Add sterile H2O to a final volume of 12 µl for each reaction.
Incubate the tubes at 65°C for 5 min
Cool the tubes on ice for 3 min
Add the following to each reaction tube:
4 µl 5X First-Strand Buffer
1 µl DTT (20 mM)
2 µl dNTP Mix (10 mM)
2 µl SmartRT reverse transcriptase
Incubate the tubes:
50°C for 45 min
85°C for 5 min

Additional information

OPTIONS

4000u(40 rxn) (100u/ul), 10000u(100 rxn) (100u/ul), 40000u(400 rxn) (100u/ul), any size

SMARTRT-REVERSE
Figure 1.Smart cDNA synthesis compared to conventional cDNA synthesis. Unlike conventional cDNA synthesis methods which involve a multiple enzyme/multiple step procedure, the Smart cDNA synthesis protocol is performed by on reverse transcription reaction, in a single tube, with no adaptor ligation or intervening purification steps. Following PCR amplification, Smart cDNA is immediately available for a variety of downstream applications.